EA031075B1

EA031075B1 - Polyguanidine derivatives as biocides

Info

Publication number: EA031075B1
Application number: EA201591381A
Authority: EA
Inventors: Александер Претч; Михаэль Нагль; Кристоф Виснер; Хайнц Бургманн
Original assignee: Силайф Фарма Гмбх
Priority date: 2013-01-25
Filing date: 2014-01-22
Publication date: 2018-11-30
Also published as: US20150368193A1; SI2948496T1; BR112015017840A2; MY184140A; JP6302487B2; PH12015501655B1; AT513858A4; CN105073835B; CA2899456C; PT2948496T; EA201591381A1; EP2948496A1; JP2016504474A; PH12015501655A1; SG11201505832SA; UA115581C2; CY1122517T1; AU2014210359A1; PL2948496T3; AU2014210359B2

Abstract

The invention relates to new polycondensation products of aminoguanidine and/or 1,3-diaminoguanidine with one or more diamines, i.e. polyguanidine derivatives of formula (I) below, wherein X is selected from -NH, aminoguanidine, and 1,3-diaminoguanidine; Y is selected from -H and -R-NH; or X and Y together stand for a chemical bond in order to produce a cyclic structure; Ris selected from divalent organic groups having 2 to 20 carbon atoms, in which one or more carbon atoms are optionally replaced by O or N; a and b are independently from each other 0 or 1, wherein a+b≠2 if no 1,3-diaminoguanidine units are present; Ris selected from -H and -NH, wherein Ris -NHif a+b=0, Ris -H or -NHif a+b=1, and Ris -H if a+b=2; and n≥2; or salts of said compound; to a production method and various applications of said products.

Description

The invention relates to the novel polycondensation products of aminoguanidine and / or 1,3 diaminoguanidine with one or more diamines, i.e. to polyguanidine derivatives of the following formula (I), where X is selected from —NH ₂ , aminoguanidine and 1,3-diaminoguanidine; Y is selected from -H and -Ri-NH ₂ ; or X and Y together form a chemical bond, forming a cyclic structure; Ri is selected from divalent organic radicals having from 2 to 20 carbon atoms, in which optionally one or more carbon atoms are substituted with O or N; a and b are independently from each other equal to 0 or 1, while a + b # 2, if the elements of 1,3-diaminoguanidine are not contained; R _{2 is} selected from -H and -NH ₂ , wherein R ₂ is -NH ₂ , if a + b = 0, R ₂ is -H or -NH ₂ , if a + b = 1, and R ₂ is is -H if a + b = 2; and n>2; or salts of said compound; to the method of production, as well as to the various applications of these products.

(I)

The invention relates to new bioactive polymers, as well as to their use as biocides.

The level of technology

Polyguanidine, having the general formula given below, as well as its various derivatives, have been known for a long time.

NH

- A, * + ^to “st n + '

In patent literature as early as 1943, US Pat. No. 2,325,586 described several methods for producing various polyguanidines by means of polycondensation of i) guanidine or its salts, ii) cyanohalide, iii) dicyanamide or iv) isocyanide dihalogenide with a diamine or v) two dicyandiamides between themselves (which gives cyanogen substituted polyguanidine), and the use of polyguanidines thus obtained as an aid in dyeing

i)

ϋ)

As diamines in reactions (i) to iv), then both alkylene- and phenylenediamines and oxyalkylene diamines or polyether diamines, which later became known as Jeffamine®, were already disclosed.

Decades later, such polyguanidines began to be used as excellent biocides. The Oskar Schmidt research group in WO 99/54291 A1 discloses the production of polyhexamethylene guanidine microbicides, in WO 01/85676 A1 polyguanidine biocides, which are obtained by the condensation of guanidine with polyoxyalkylene, and in WO 2006/047800 A1, which act as biocides, in particular as fungicides of polyanaline, and in WO 2006/047800 A1, which act as biocides, in particular as fungicides of polyanaline, and in WO 2006/047800 A1, which act as biocides, in particular as fungicides of polyanaline, and in WO 2006/047800 A1. formed during the polycondensation of guanidine with a mixture of alkylenediamine and oxoalkylenediamine and should have a lower toxicity than polymers that contain only one of the two types of divalent radicals R _b

WO 02/30877 A1 describes a similar polyguanidine as a disinfectant, which additionally contains phenylene groups in the chain. A group of Russian researchers (Tets, Tets and Krasnov) in WO 2011/043690 A1, on the basis of which US 2011/0269936 A1 and EP 2520605 A1 were submitted, disclose polyguanidine biocides of the following formula, which are obtained by polycondensation of guanidine and hexamethylenediamine in the presence of hydrazine hydrate

Hydrazine during polycondensation, at least formally, replaces the amino group in either only one or two guanidine groups, as a result of which block copolymers should be formed, in which poly (hexamethylene-guanidine) blocks alternate with poly (hexamethylaminoguanidine) blocks,

- 1 031075 and both types of blocks are connected to each other through guanidine dimers, as shown below

Also, these polymers and their acid addition salts should act as biocides against bacteria, viruses, and fungi. However, in the examples of this application, in which 7 different polymers were obtained, no physical data of the products obtained were indicated, except as indicated in Example 1: a solid, almost colorless, transparent substance.

Regarding the possible structure of the compound that can be formed during the polycondensation of guanidine with diamine, there are several articles by the research group of the Technical University of Graz, for example Albert et al., Biomacromolecules 4 (6), 1811-1817 (2003) and Feiertag et al., Macromol . Rap. Comm. 24 (9), 567-570 (2003). In addition to the various possibilities of forming linear polymer chains from the corresponding starting monomers, they are usually formed in a non-negligibly small amount, which among other things depends on the length of the diamine chain, are also cyclic molecules of the following general formula

The main disadvantages of almost all the polyguanidine derivatives described above are, on the one hand, in the non-negligible toxicity of these products, as well as in the case of the use of highly reactive components in a relatively expensive production method, as well as in toxicological components, such as hydrazine, with the result that the invention is to obtain new, less toxic, but still effective as biocides of polyguanidines as simple as possible m and economical way, avoiding the drawbacks mentioned above.

DISCLOSURE OF INVENTION

The specified purpose in this invention is achieved by the provision of new polycondensation products of aminoguanidine and / or 1,3-diaminoguanidine with one or more diamines, which are polyguanidine derivatives of the following formula (I)

(I) where X is selected from —NH ₂ , aminoguanidine and 1,3-diaminoguanidine;

Y is selected from —H and —R ₁ —NH ₂ ;

or X and Y together form a chemical bond, forming a cyclic structure;

R1 is selected from alkylene radicals having from 2 to 20 carbon atoms, in which optionally one or more carbon atoms are substituted with O or N;

a and b are independently from each other equal to 0 or 1, while a + b Φ 2, if the elements of 1,3-diaminoguanidine are not contained;

R2 is selected from -H and -NH2, wherein R ₂ is -NH ₂ , if a + b = 0,

R ₂ is -H or -NH ₂ if a + b = 1 and

R ₂ is -H if a + b = 2;

and n>2;

or salts of the compounds.

The new derivatives of polyguanidine of formula (I) in tests for activity have shown themselves to be effective antimicrobial substances, which, however, unexpectedly have significantly less toxicity than polymers with a similar structure from the above-cited documents WO 2011/043690 A1, US 2011/0269936 A1 and EP 2520605 A1, which is confirmed by the later examples of the implementation of this invention and comparative examples. Without intending to be limited to one theory, the inventors suggest that amino groups and diaminoguanidine groups are better compatible with human eukaryotic cells than guanidine groups and, in particular, those polymers that contain the hydrazine bridges of guanidine dimers described above. In addition, in the disclosed method, the use of the toxic component of hydrazine hydrate is avoided in the polymerization process, which in some polymers according to the prior art may be contained as residual monomer.

The compound of the above formula (I) is represented as a product of polycondensation (Mo

- 2 031075 but) aminoguanidine, hereinafter referred to as MAG, and the product of polycondensation and 1,3 diaminoguanidine, hereinafter referred to as DAG

Formula (I) is explained by the fact that during the polycondensation proceeding with the removal of ammonia MAG and DAG radicals can take part in the polycondensation both with their amino or tautomeric imino groups and with their hydrazo (hydrazinyl) groups. Consequently, for both MAG and DAG, as a starting monomer, there are three different possibilities for incorporation into the polymer chain of the invention. In the case of MAG in formula (I), the only hydrazo group, and in the case of DAG, on the contrary, a single imino / amino group can be directed to the right, left, or upward.

This means for MAG the following parameters in formula (I):

a = 1, b = 0, R ₂ is -H: the hydrazogroup is directed to the left;

a = 0, b = 1, R ₂ is -H: the hydrazogroup is directed to the right; or a = 0, b = 0, R ₂ is -NH ₂ : the hydrazogroup is directed upward.

For DAG, the following options are available for parameter combinations:

a = 0, b = 1, R2 is -NH2: the amino / imino group is directed to the left;

a = 1, b = 0, R ₂ is —NH ₂ : the amino / imino group is directed to the right; or a = 1, b = 1, R ₂ is -H: amino / imino is directed upwards.

Not limited to this, the NMR spectra of the polycondensates obtained, as described later for the present invention, confirm that during the polycondensation reaction mixtures of some of the three possible orientations are almost always formed, and it must be assumed from this (it turned out not 100%). that even within the same chain there are different orientations of the same monomer.

In connection with the above, it should be made clear that the position of the double bond C = N guanidino, as well as the spatial position of R ₂ in the double bond, is usually influenced by tautomerism. This means that the guanidine double bond can be located inside the chain or outside the chain, and R ₂ can be directed to the left or right. Therefore, such tautomers of the above-described polycondensation products of formula (I) should also be within the protection scope of this invention.

The above options for X and Y are obtained due to the different possibilities of chain formation depending on whether MAG, DAG or a mixture of both of the original monomers are used, including the possibility of loop closure in a cyclic polycondensation product. See also pre-cited articles by Albert et al. and Feiertag et al. For terminal aminoguanidino- (MAG-) and 1,3-diaminoguanidino- (DAG-) groups there are of course the same possibilities as for those that are inside the chain, i.e. chaining can occur through any nitrogen atom.

The radical R ₁ according to this invention may be a linear, branched or cyclic, saturated or unsaturated, divalent hydrocarbon radical having from 2 to 20 carbon atoms, preferably from 4 to 18 carbon atoms, more preferably from 6 to 12 carbon atoms, in which some C atoms may be replaced by O and / or N. The above preferences were obtained from the following considerations. With a very short radical R _{1, the} active MAG or DAG groups are very close to each other, which can reduce the activity of the polymer; in the case of a long radical, the active groups on the contrary are too far apart. Therefore, radicals with more than 20 atoms can be fundamentally taken into account, but from an economic point of view they do not look preferable, since they provide polymers of formula (I), in which a relatively small number of guanidino groups with anti-infective action are contained per unit mass.

Preferably, the radical R ₁ is selected from alkylene radicals in which optionally one or more carbon atoms are replaced by O or N, which increases the hydrophilicity of the chain, even more preferably selected from radicals with the following general formulas (II) to (V) - (CH _₂₎ —— Z— (CH ₂ ) _d - (I) - (CH ₂ ) _s —z— (CH ₂ ) _d - z ₂ - (CH ₂ ) _e - (in) - (C H ₂ ) _s —Z— (C H ₂ ) _d - z ₂ - (CH ₂ ) _e — z ₃ - (CH ₂ ) _f - (iv) (CH ₂ ) _with Z ₁ (CH ₂ ) _d Z ₂ (CH ₂ ) _e Z ₃ ( CH ₂ ) _f Z ₄ (CH ₂ ) _g (V)

- 3 031075 where from Z ₁ to Z ₄ independently of one another represent a heteroatom selected from O and N, and the indices c from g to g independently of each other are an integer from 1 to 12, that the total number of atoms of the radical R ₁ does not exceed 20. It is especially preferable that inside the radical R ₁ all Z heteroatoms are either O or N.

The best results when testing biocidal action or toxicity were achieved with those compounds in which R ₁ was a bivalent radical of a simple polydiamine, such as, for example, 4,9-dioxadodecane-1,12-diamine, polyoxyethylene and / or propylene diamine, with In this case, n was preferably equal to from 2 to 15, more preferably from 2 to 10, in particular from 2 to 6.

As salts of new polyguanidines of formula (I), any acid addition salt with one or several inorganic or organic acids, such as, for example, hydrohalic acids, oxygen acids of nitrogen, sulfur or phosphorus, boric acid, carbonic acid, carboxylic acids, thiocarboxy-, carbamic-, sulfonic-, phosphonic- or phosphinic acids, as well as partial esters or amides of multivalent representatives of these acids. Pharmaceutically acceptable salts are preferred according to the invention, more preferably acid addition salts, such as hydrochloride, hydrobromide, hydroiodide, sulfate, methyl sulfate, carbonate, borate, cyanate, thiocyanate, phosphate, mesylate, nitrate, acetate, benzoate, lactate, tartrate, citrate , maleate, fumarate or partial esters of these acids, if they are two or more functional. It is preferable to use pharmaceutically acceptable alcohols, in particular ethanol, as the alcohol component of such partial esters.

If Ri contains one or more OH or COOH groups, then a salt with an inorganic or organic base also falls within the protection scope of the present invention, with salts with a pharmaceutically acceptable base, more preferably with a guanidine derivative, in particular with an amino or diaminoguanidine, i.e. with a guanidine derivative, from which new polyguanidines according to the invention are produced. As a rule, upon receipt, internal salts are formed from acidic and basic groups within the corresponding molecule.

In a second aspect of the present invention, there is provided a method for producing polyguanidine derivatives according to the invention according to a first aspect by polycondensation of guanidine derivatives or their salts with a diamine, which is characterized in that MAG and / or DAG or their acid addition salt is polycondensed with at least one diamine H ₂ NR ₁ —NH ₂ when heated.

In contrast to the prior art, the method of the invention involves the reaction of a MAG or DAG with one or more diamines, preferably with only one diamine. As a result, it is possible to obtain more clearly defined products than in the works of Russian researchers cited above, especially since free hydrazine was not detected in any reaction phase of the reaction mixture obtained according to the invention, neither chromatographically nor chemically. Adverse reactions with hydrazine, which were desirable in a certain prior art, are completely undesirable in this case and can thus be effectively avoided.

Preferably, the method according to the invention is carried out in such a way that the salt MAG or DAG, in particular the hydrochloride salt, together with the diamine, which is preferably used in a slight molar excess, for example from 3 to 5 mol.%, Because of the economy of a maximum of 10 mol.%, Relative to to (di) aminoguanidine in order to ensure complete conversion of the guanidine derivative, it is first heated to a first, lower temperature, preferably to about 80-150 ° C, more preferably to 110-130 ° C, and then to a second, higher temperature, preferred It is 15050 ° C, more preferably 160-180 ° C, in order to increase the reaction rate and with it the evolution of gas. While the reaction mixture is preferably from 1 to 3 hours, more preferably about 2 hours, kept at the first temperature and then preferably from 1 to 8 hours, more preferably from 3 to 5 hours, kept at the second temperature in order to ensure complete passage of the reaction.

The reaction is preferably carried out at normal pressure as well as with the removal of water, which is caused, for example, by at least the initial purging of the reactor with an inert gas and equipping the reactor with a drying tube. However, it is also possible to apply a vacuum, especially at the end of the reaction, in the form of a purification stage in order to completely remove free ammonia, as well as residual monomers, i.e. first of all excess diamine.

After completion of the reaction, the resulting polyguanidine derivative is preferably dissolved in water, for example, in about 3 to 10 times the amount of water. On the one hand, this allows the separation of possible water-insoluble components, and on the other hand, an aqueous solution is the preferred form of release for the application of new polymers, i.e. they can be directly applied as a solution, optionally after the addition of adjuvants.

Another, currently less preferred purification option is, for example, evaporation of water from an aqueous solution and drying the polymer under vacuum, or salting out of an aqueous solution by adding acid followed by drying, which takes into account the above-mentioned acids, in particular, described as preferred pharmaceutically acceptable acid. One

- 4 031075 embodiment of salting out includes the introduction of CO ₂ and salting out of polyguanidine as carbonate or bicarbonate. If the desired polyguanidine needs to be applied not as a salt, but as a free base, treatment with a base, which is in the form of an aqueous or non-aqueous solution or suspension, can follow salting out.

In the third aspect, the present invention relates to the use of a polyguanidine derivative according to the first aspect of the present invention for combating bacterial, fungal and viral infections and their effects, as a pesticide and disinfectant, and mainly as a disinfection agent (biocide) for reducing the number and killing microorganisms . Further, the invention is described in more detail using non-limiting examples of implementation, together with comparative examples. The figure shows the results of toxicity tests.

Examples

Examples 1-6 and comparative examples 1 and 2. The preparation of the polymer.

Example 1

1,3-diaminoguanidine hydrochloride mmol and 4,9-dioxadodecane-1,12-diamine hydrochloride were heated in a reactor, which was closed with a drying tube, for 90 minutes with stirring to 120 ° C, then the temperature was increased to 100 ° C for 100 minutes, from this time at the end of the reaction 45 min. was kept under reduced pressure (50 mbar). After cooling the reaction mixture to a temperature below 80 ° C, 25 ml of water was added to the gel-like reaction product. After several hours, a clear solution was obtained.

From the sample of the resulting aqueous solution, water was evaporated, and the resulting precipitate was dried in vacuo to give a reddish, viscous liquid. This liquid was dissolved in 2 ml of D ₂ O (with a degree of deuteration of> 99.5%), and the 'H-NMR spectrum was measured. The position of the thus distinguished groups of protons of methylene in the radical Ri in the product was as follows:

¹ H-NMR (D ₂ O), δ (ppm): 1.54-1.67 (m, OCH ₂ CH ₂ CH ₂ CH ₂ O), 1.80-1.95 (m, NCH ₂ CH ₂ ), 3.23-3.38 ppm (m, NCH ₂ ), 3.42-3.65 ppm (m, CH ₂ CH ₂ OCH ₂ CH ₂ ).

This confirmed the structure of the used component of the diamine, 4,9-dioxadodecane-1,12-diamine. Example 2

4.6 mmol of 1,3-diaminoguanidine hydrochloride and 4.8 mmol of 4.9-dioxadodecane-1.12-diamine were heated in a reactor, which was closed with a drying tube, for 90 minutes with stirring to 120 ° C, then the temperature was raised for 8 hours to 180 ° C, from this time at the end of the reaction 45 minutes was kept under reduced pressure (50 mbar). After cooling the reaction mixture to a temperature below 80 ° C, 16 ml of water was added to the gel-like reaction product. After several hours, a clear solution was obtained.

Example 3

4.6 mmol of N-aminoguanidine hydrochloride and 4.8 mmol of 4.9-dioxadodecane-1.12-diamine were heated in a reactor, closed with a drying tube, for 90 minutes with stirring to 120 ° C, then the temperature was raised for 3.5 hours to 180 ° C, from this time at the end of the reaction 60 min was kept under reduced pressure (50 mbar). After cooling the reaction mixture to a temperature below 80 ° C, 16 ml of water was added to the gel-like reaction product. After several hours, a clear solution was obtained.

Example 4

1.16 mmol of 1,3-diaminoguanidine hydrochloride and 1.21 mmol of tris (2-aminoethyl) amine were heated in a reactor closed in a drying tube for 150 minutes with stirring to 120 ° C, then the temperature was raised to 2.5 hours within 2.5 hours ° C, from this time at the end of the reaction 45 min was kept under reduced pressure (50 mbar). After cooling the reaction mixture to a temperature below 80 ° C, 4 ml of water was added to the gel-like reaction product. After several hours, a clear solution was obtained.

Example 5

8.12 mmol of 1,3-diaminoguanidine hydrochloride and 8.47 mmol of tris (2-aminoethyl) amine were heated in a reactor closed in a drying tube for 120 minutes with stirring to 130 ° C, then the temperature was raised to 180 ° C for 8 hours From this time, at the end of the reaction for 90 minutes, it was kept under reduced pressure (15 mbar). After cooling the reaction mixture to a temperature below 80 ° C, 28 ml of water was added to the gel-like reaction product. After several hours, a clear solution was obtained.

Example 6

2.32 mmol of 1,3-diaminoguanidine hydrochloride and 2.43 mmol 3, b-dioxaoctane-1,8-diamine was heated in a reactor, which was closed with a drying tube, for 60 minutes with stirring to 120 ° C, then the temperature was raised for 4 hours to 170 ° C, from this time at the end of the reaction 60 min was kept under reduced pressure (50 mbar). After cooling the reaction mixture to a temperature below 80 ° C, 7 ml of water was added to the gel-like reaction product. After several hours, a clear solution was obtained.

- 5 031075

Comparative example 1.

23.2 mmol of guanidine hydrochloride, 5.4 mmol of 3,6-dioxaoctane-1,8-diamine and 18.1 mmol of 1,6 diaminohexane were heated in a reactor, closed with a drying tube, for 90 minutes with stirring to 120 ° C, then the temperature in for 8 h, raised to 170 ° C, from this time at the end of the reaction 90 min was kept under reduced pressure (50 mbar). After cooling the reaction mixture to a temperature below 80 ° C, 60 ml of water was added to the gel-like reaction product. After several hours, a clear solution was obtained.

The polymer obtained was structurally consistent with that disclosed in WO 2006/047800 A1.

Comparative example 2.

2.00 mmol of guanidine hydrochloride, 1.70 mmol of 1,6-hexamethylenediamine and 0.3 mmol of hydrazine hydrate were heated in a reactor, which was closed with a drying tube, for 90 minutes with stirring to 160 ° C, then the temperature was raised for 3.5 hours to 180 ° C, from this time at the end of the reaction 60 min was kept under reduced pressure (50 mbar). After cooling the reaction mixture to a temperature below 80 ° C, 4 ml of water was added to the gel-like reaction product. After several hours, a clear solution was obtained.

The polymer obtained was structurally consistent with that disclosed in WO 2011/043690 A1.

Example 7. Determination of antimicrobial / antifungal / antiviral activity.

The activity of the new compounds was tested in repeated tests in screening systems. Antibacterial and antifungal activity was investigated using OLS-test. MNC is the minimum inhibitory concentration (English: MIC fur minimal inhibitory concentration) and means the lowest concentration of a substance at which no multiplication of microorganisms is detected by the naked eye. OLS is determined by the so-called titration method, in which the substance is diluted and then the pathogen is added. As a rule, the concentration of the antibiotic is determined in this way, which still slows the growth of bacterial strains. OLS is indicated in micrograms per milliliter (μg / ml) or in vol.%, And dilution usually occurs on a log2 scale. In this case, the initial concentration of 1% was diluted twice each time, which gave test concentrations of 0.5, 0.25, 0.125%, etc. Thus, the lowest value reflects the best activity as an anti-infective agent.

The tests were carried out in accordance with the standards of EUCAST (European Committee for Antimicrobial Susceptibility Testing (European Committee for the Testing of Antimicrobial Sensitivity)) and according to the instructions of the AFST (Antifungal Susceptibility Testing) European Society of Clinical Microbiology and Infectious Diseases (ESCMID).

A screening system for viruses is an infection system in which host cells are infected under laboratory conditions, and the test substance is added before or after infection, and its activity is determined. All of these tests were performed according to SeaLife Pharma's internal standard drug screening guidelines, with similar dilution series used for antibacterial / antifungal testing.

The following table. 1-3 shows the results of tests on the anti-infective effect of the new compounds according to the invention of examples 1, 3-5 against some multi-resistant bacteria and fungi, as well as viruses. The data given are average values of multiple measurements.

It is obvious that the new compounds according to the invention demonstrate excellent activity against both gram-positive and gram-negative pathogens.

Table 1

OLS test result Mrsa Staphylococcus epidermis Streptococcus pneumoniae Enterococcus faecalis Propioni bacter acne E. coli Klebsiella pneumoniae Pseudominas aeruginosa Acinetobacter baumanii Enterobacter cloace Salmonella enterica Example 1 0.001% 0.001% 0.004% 0.008% 0.001% 0.016% 0.02% 0.02% 0.06% 0, 03% 0, 03% Example 3 0.001% 0.001% 0.001% 0.008% 0.001% 0.02% 0.02% 0.02% 0.06% 0.2% 0, 03% Example 4 0.001% 0.001% 0.001% 0.008% 0.001% 0.016% 0.016% 0.030% 0.02% 0.016% 0.030% Example 5 0.001% 0.001% 0.002% 0.002% 0.001% 0.020% 0.02% 0, 04% 0, 04% 0, 13% 0, 03%

And also against fungi and yeast.

- 6 031075

table 2

OLS test result Candida albicans Candida papillosis Candida glabrata Candida kruzei Aspergillus terreus Aspergillus fumigatus Fusarium rose! Trichophyton sp. Alternarria alternarria Microsporum canis Dematiacea sp. Example 1 0.008% 0, 03% 0.02% 0.02% 0.02% 0, 03% 0, 03% 0.02% 0.02% 0, 03% 0.02% Example 3 0.02% 0.02% 0.02% 0.02% 0, 03% 0, 03% 0, 03% 0.02% 0.02% 0.02% 0.02% Example 4 0.008% 0.016% 0.016% 0.008% 0.125% 0.125% n. t. nt nt nt nt Example 5 0.02% 0.02% 0.02% 0.020% 0.016% 0.016% nt nt nt nt nt

And also against viruses.

Table 3

The results of the virological test Influenza A and B Human rhinovirus Parainfluenza virus Herpes simplex virus Example 1 0.008% 0.008% 0.008% 0.02% Example 3 0.02% 0.02% 0.02% 0.02% Example 4 0, 04% 0.02% 0, 04% 0.02% Example 5 0, 04% 0.04% 0, 04% 0.02%

Thus, all tested new compounds have activity from very good to excellent against a wide variety of pathogens and with significantly less toxicity than those of polyguanidine derivatives known from the prior art, as shown by the following toxicity tests.

Example 8. Tests for toxicity.

Using the AlamarBlue® assay, as described below, 4 polymers analyzed toxicological potential (including proliferation, cell death, cell metabolism) and determined the IC ₅₀ value, as well as the non-toxic concentration in primary keratinocytes (HKER) and primary endothelium cells (HUVEC). The figure shows the toxic effect of various polymers depending on the concentration.

AlamarBlue® analysis: 20,000 human keratinocytes (HKER) or endothelial cells (HUVEC) were placed in a 96-well plate and incubated for 24 hours before new polymers from examples 1 and 3 were added in different concentrations (from 5 to 0.005%), and also comparative substances from comparative examples 1 and 2. After 24 hours, 10 μl of AlamarBlue® was added to each well (100 μl of medium) and after 3 hours of incubation, the stained reaction was evaluated using a plate reader spectrophotometer (excitation: 530 nm; emission: 590 nm). HKER: human primary keratinocytes, human primary keratinocytes; HUVEC: human umbilical vein endothelial cells, human endothelial cells of the umbilical cord vein.

The polymers of comparative examples 1 and 2 have both for HKER and HUVEC already at very low concentrations, i.e. IC50 is about 0.01% or lower, with a significant toxic effect. The new polymers of examples 1 and 3 obtained by the inventors showed a toxic effect only at significantly higher concentrations compared with comparative examples: the IC50 is for example 1 for both cell types in the 1% area and for example 3 in the range from 0.05 to 0, one%. The toxicity of comparative examples is achieved by the polymer from example 3 only at at least 5 times the concentration, and with the polymer from example 1 only at at least 100 times the concentration. Thus, in this test, the derivative DAG showed a clearly better result than the polymer MAG.

Thus, the new compounds have a very good to excellent activity against a wide variety of pathogens with a significantly lower toxicity than the polyguanidine derivatives known from the prior art.

Claims

CLAIM

1. The product of the polycondensation of aminoguanidine and / or 1,3-diaminoguanidine with one or more diamines, which is a polyguanidine derivative of the following formula (I):

where X is selected from —NH ₂ , aminoguanidine and 1,3-diaminoguanidine;

- 7 031075

Y is selected from -H and -R _r NH ₂ ;

or X and Y together form a chemical bond, forming a cyclic structure;

R _{1 is} selected from alkylene radicals having from 2 to 20 carbon atoms, in which optionally one or several carbon atoms are replaced by O or N;

a and b are independently from each other equal to 0 or 1, while a + b Ψ 2, if the elements of 1,3-diaminoguanidine are not contained;

R _{2 is} selected from -H and -NH ₂ , wherein R ₂ is -NH ₂ if a + b = 0,

R ₂ is -H or -NH ₂ if a + b = 1 and

R ₂ is -H if a + b = 2;

and n> 2;

or a salt of the compound indicated.

2. The polyguanidine derivative according to claim 1, wherein R _{1 is} selected from radicals with the following general formulas from (II) to (V):

--- (CH ₂ ) _s --- 7 - (CH ₂ ) _d - (I)

- (CH ₂ ) _s --- 7 - (CH ₂ ) _d --- Z ₂ - (CH ₂ ) _e - (III) - (CH ₂ ) _c - 7— (CH ₂ ) _d - z ₂ - (CH ₂ ) _e — z ₃ - (CH ₂ ) _f - (IV) - (CH ₂ ) _c - 7 - (CH ₂ ) _d - z ₂ - (CH ₂ ) _e — z ₃ - (CH ₂ ) _f —z ₄ - (CH ₂ ) _g - (V) r

where from Z ₁ to Z ₄ independently of one another represent a heteroatom selected from O and N, and the indices c to g independently of each other are such an integer from 1 to 12 that the total number of atoms of the radical R ₁ does not exceed 20.

3. Derived polyguanidine according to claim 2, characterized in that inside the radical R ₁ all Z heteroatoms are either O or N.

4. The polyguanidine derivative according to claim 3, wherein R ₁ is a divalent polyether diamine radical.

5. Derived polyguanidine in one of the paragraphs.1-4, characterized in that n = from 2 to 6.

6. Derived polyguanidine in one of the paragraphs.1-5, characterized in that it is an acid addition salt in the form of hydrochloride, hydrobromide, hydroiodide, sulfate, carbonate, borate, cyanate, thiocyanate, phosphate, mesylate, nitrate, acetate, benzoate , lactate, tartrate, citrate, maleate, fumarate, or a mixture of two or more of these salts.

7. A method of producing a polyguanidine derivative according to one of claims 1 to 6 by polycondensation of a guanidine derivative or its salt with a diamine, characterized in that aminoguanidine or 1,3 diaminoguanidine or its acid addition salt is subjected to polycondensation with at least one diamine H ₂ NR ₁ -NH ₂ when heated, where R ₁ is as defined in claim 1.

8. The method according to claim 7, characterized in that at least one diamine is used in an excess of from 3 to 5 mol.% In relation to the guanidine derivative.

9. The method according to claim 7 or 8, characterized in that the salt of aminoguanidine or 1,3-diaminoguanidine together with at least one diamine is first heated to the first, lower temperature, and then to the second, higher temperature.

10. The method according to claim 9, characterized in that the salt of aminoguanidine or 1,3-diaminoguanidine together with at least one diamine is first heated to 110-130 ° C, and then to 160-180 ° C.

11. The method according to p. 9 or 10, characterized in that the reaction mixture is maintained from 1 to 3 hours at the first temperature and from 1 to 8 hours at the second temperature.

12. The method according to one of claims 7 to 11, characterized in that the obtained polyguanidine derivative is purified by dissolving in from about 3 to about 10 times the amount of water.

13. Use of a polyguanidine derivative according to one of claims 1 to 6 for combating bacterial, fungal and viral infections and their consequences.

14. The use of a derivative of polyguanidine in one of claims 1 to 6 as a pesticide and disinfection agent.

15. The use according to claim 14 for disinfection as a disinfectant (biocide) to reduce the number and kill microorganisms.

16. The use of a polyguanidine derivative according to one of claims 1 to 6 for use according to any of claims 13 to 15, characterized in that the effective amount of the polyguanidine derivative is in the form of a solution in from 3 to 10 times the amount of water.

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